Saverio Brogna (Genetics) (sb at mole.bio.cam.ac.uk) wrote:
: Dear someone,
: I have expressed e protein in E. coli with differents systems (His-tag,
: Glutation-transferase fusion and wild-type) but I can not get it soluble.
: DO YOU KNOW HOW TO EXPRESS PROTEINS IN A WAY THAT SOME IS SOLUBLE?
: Thanks
: --
: Saverio Brogna
: University of Cambridge, Departement of Genetics
: Downing Street, Cambridge, CB2 3EH, England (UK)
: tel +44-223-333970, fax +44-223-333992
You might like to try one of the systems that direct expression to the
periplasm. I have used pASK-60 from Biometra which worked OK. Then there's
pET12a which should do the same.
Alternatively you could have a go at refolding it, but it can take a while
to get the conditions right.
Debbie
--
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Debbie Hodges dh124 at cus.cam.ac.uk Fax 01223 217838
Rheumatology Research Unit, Addenbrookes Hospital, Hills Road, Cambridge, UK.
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